-
00:15
1.
Comparative Gene-expression Analysis
-
00:24
2.
Question
-
00:39
3.
Differential Display
-
02:55
4.
Polymerase Chain Reaction
-
03:09
5.
Template DNA
-
01:32
6.
SpecificityMust be complementary to flanking sequences of target regionNot be complementary to other non-target regions of genomeNested polymerase chain reaction (nested PCR): to reduce non-specific binding Primer LengthToo short lack specificityToo lon
-
02:38
7.
Product SizeBasic Taq polymerase can easily amplify fragments up to 1,000 to 2,000 bpTypical qPCR amplicons are 70-200 bp in lengthToo long Fluorescent intensity not in linear rangePrimer DimersIf the primers have self-complementary sequences, the prime
-
05:10
8.
General Considerations in Primer Design
-
01:16
9.
General Considerations in Primer Design
-
00:42
10.
G/C ContentPrimers should be about 50% G/C (40~60%) Not have long runs of G/C or A/T (>3 bp) A stretch of A/T’s might only weakly base pairA stretch of G/C might promote mis-annealingG/C clampTo ensure the stability of this interaction, primers are often
-
00:07
11.
Web Based Tools for Primer Design
-
00:09
12.
http://biotools.nubic.northwestern.edu/OligoCalc.html
-
00:05
13.
Web Based Tools for Primer Design
-
00:05
14.
https://www.ncbi.nlm.nih.gov/tools/primer-blast/
-
01:09
15.
Question
-
01:27
16.
Rapid Amplification of cDNA Ends (RACE)
-
01:00
17.
Rapid Amplification of cDNA Ends (RACE)
-
02:13
18.
Multiplex PCR
-
00:08
19.
Primer Design Assistant
-
03:28
20.
PCR-based Techniques: Subtractive hybridization
-
00:58
21.
QuestionCan RNA be amplified directly by PCR? Why and why not?
-
02:00
22.
Reverse Transcription PCR
-
03:05
23.
Real Time - PCR
-
02:41
24.
Real Time - PCR
-
04:27
25.
Amplification Plot